Chitosan derivatives killed bacteria by disrupting the outer and inner membrane

Six kinds of water-soluble chitosan were prepared by grafting aminofunctionality onto chitosan at the C-6 position, and their antimicrobial activities were investigated against three Gram-negative and three Gram-positive bacteria. Among the derivatives, dimethylaminoethyl-chitosan (DMAEC) has the highest potential to suppress the growth of bacteria. To elucidate detailed antimicrobial modes of action against bacteria, cell integrity, outer membrane (OM), and inner membrane (IM) permeabilization assays were investigated. When treated with DMAEC, the release of 260 nm absorbing materials quickly increased for both Escherichia coli and Staphylococcus aureus, but the absorbance value was different due to the difference in cell structures. In OM and IM permeabilization assays, DMAEC rapidly increased 1-N-phenylnaphthylamine uptake and the release of cytoplasmic beta-galactosidase via an increase in the permeability of OM and IM. Moreover, DMAEC90 prepared from 90% deacetylated chitosan had more activity than DMAEC50 prepared from 50% deacetylated chitosan, and these results revealed that the antimicrobial action of water-soluble chitosans was dependent on the degree of deacetylation and the substituted group.     

A novel angiotensin I converting enzyme inhibitory peptide from Alaska pollack (Theragra chalcogramma) frame protein hydrolysate

Alaska pollack frame protein, which is normally discarded as an industrial byproduct in the processing of fish in plants, was hydrolyzed with pepsin. This was fractionated into five major types of Alaska pollack frame protein hydrolysates (APH-I, 10-30 kDa; APH-II, 5-10 kDa; APH-III, 3-5 kDa; APH-IV, 1-3 kDa; and APH-V, below 1 kDa) using an ultrafiltration membrane bioreactor system. Angiotensin I converting enzyme (ACE) inhibitory activities of the fractionated hydrolysates were investigated, and the fraction that exhibited the highest ACE inhibitory activity was further purified using consecutive chromatographic methods on SP-Sephadex C-25 column, Sephadex G-25 column, and high-performance liquid chromatography (HPLC) on an octadecylsilane column. Finally, we purified a novel ACE inhibitory peptide with an IC50 value of 14.7 microM, and the sequence of the peptide was Phe-Gly-Ala-Ser-Thr-Arg-Gly-Ala. In addition, the ACE inhibition pattern of the peptide was found to be noncompetitive.     

Ochratoxin A in Korean food commodities: occurrence and safety evaluation

To evaluate the exposure of Koreans to ochratoxin A, we conducted a survey in 2003 for ochratoxin A in various domestic food commodities: 60 polished rices, 22 barleys, 35 wheat flours, 46 beers, and 14 unstrained rice wine (makkolli) samples. They were analyzed for ochratoxin A using immunoaffinity column and high-performance liquid chromatography (HPLC)-fluorescence detection, and the positive samples were confirmed using HPLC-tandem mass spectrometry. By combining results from different surveys on the levels of ochratoxin A in selected foods and the consumption patterns, we obtained the Korean probable daily intakes (PDI) of ochratoxin A. The polished rice commodity had the highest mean levels of ochratoxin A, which ranged from 0.2 (not detected, i.e., ND = 0) to 1.0 ng/g (ND = limit of detection, i.e., LOD). The estimated PDI for all Koreans fell into the range of 0.8-4.1 ng/kg bw/day, while for heavy consumers the estimates ranged from 1.7 to 9.1 ng/kg bw/day, which did not exceed the PTDI value (14 ng/kg bw/day). Staple rice is the major contributor (>90%) to the Korean dietary intake of ochratoxin A. On the basis of these estimates, it may be concluded that there is at present no considerable risk of ochratoxin A exposure for the average Korean consumer.     

Allelic relationships at the pvr1 locus in Capsicum annuum

Diseases caused by members of the Potyviridae family currently threaten pepper crops (Capsicum annuum L.) worldwide. A series of monogenic recessive resistance genes that control potyvirus resistance at the pvr1 locus in Capsicum species are widely known and used in pepper breeding programs, each allele with a differential resistance spectrum affecting a distinct range of viral strains across three viruses, Tobacco etch virus (TEV), Pepper mottle virus and Potato virus Y. In this study, we systematically analyzed the resistant spectra, and the level of the resistance each allele confers using a set of pepper genotypes homozygous or heterozygous for the following alleles; Pvr1 ⁺, pvr1, pvr1 ¹ and pvr1 ². The resistance alleles at the pvr1 locus show recessive inheritance when combined with a susceptible allele in F1 progenies. However, our results show that resistance in this system is, in fact, not always fully recessive and establish a hierarchy of allelic interactions We identified the resistance phenotype in F₁ progenies generated by combining a complete resistant allele with an incomplete resistance allele against TEV strains implying that the resistance alleles at the pvr1 shows dominant inheritance when combined with an incomplete resistance allele. Resistance alleles against TEV-HAT, pvr1 and pvr1 ², show dominance inheritance when each is combined with pvr1 ¹. The resistance allele, pvr1, shows dominance inheritance when combined with pvr1 ² against TEV-N. These results clarify the allelic relationship between resistance alleles at the pvr1 locus displaying different resistance spectra, and will assist breeders to select the preferred combinations of the resistance alleles to obtain durable resistance against multiple potyviral strains.     

Isolation and Characterization of Efficient Active Compounds Using High-Performance Centrifugal Partition Chromatography (CPC) from Anti-Inflammatory Activity Fraction of Ecklonia maxima in South Africa

Ecklonia maxima is a brown seaweed, which is abundantly distributed in South Africa. This study investigated an efficient approach using high-performance centrifugal partition chromatography (HPCPC), which has been successfully developed for the isolation and purification of phlorotannins, eckmaxol, and dieckol from the ethyl acetate fraction of E. maxima (EEM). We evaluated EEM for its inhibitory effect against lipopolysaccharide (LPS)-induced inflammatory responses in zebrafish embryos. The separation of eckmaxol and dieckol from samples of EEM using HPCPC was found to be of high purity and yield under an optimal solvent system composed of n-hexane:ethyl acetate:methanol:water (2:7:3:7, v/v/v/v). To evaluate the anti-inflammatory efficacy of EEM containing active compounds, zebrafish embryos exposed to LPS were compared with and without EEM treatment for nitric oxide (NO) production, reactive oxygen species (ROS) generation, and cell death two days after fertilization. These evaluations indicate that EEM alleviated inflammation by inhibiting cell death, ROS, and NO generation induced by LPS treatment. According to these results, eckmaxol and dieckol isolated from brown seaweed E. maxima could be considered effective anti-inflammatory agents as pharmaceutical and functional food ingredients.     

Experimental infection of normal and immunosuppressed pigs with Pseudomonas pseudomallei

A single dose of 5 x 10(8) bacilli of Pseudomonas pseudomallei by intratracheal injection resulted in acute (21 cases) or chronic (19 cases) melioidosis in 40 of 48 pigs. Fifteen (10 acute and 5 chronic) had been immunosuppressed by cyclophosphamide before inoculation. The major clinical signs were initial fever, marked neutrophilia and, in the acute cases, respiratory distress. There were no signs of the nasal and ocular discharge, paresis or diarrhoea seen in acute cases in south-east Asia. The cyclophosphamide treatment caused a significant decrease in the neutrophil count by 7 d after inoculation in all 15 immunosuppressed pigs, and all were culture positive at necropsy. Eight of the 33 non-treated pigs were culture negative at necropsy. Pigs overcoming the initial phase of infection had more abscess-like nodules that were bacteriologically sterile at necropsy than the pigs with acute cases of melioidosis. P. pseudomallei was isolated predominantly from the spleen, lungs and the injection site. Although only one strain was used in this study, it is likely that Australian strains of P. pseudomallei are not as virulent as the south-east Asian isolates.     

Therapeutic Potential of Phlorotannin-Rich Ecklonia cava Extract on Methylglyoxal-Induced Diabetic Nephropathy in In Vitro Model

Advanced glycation end-products (AGEs) play a vital role in the pathogenesis of diabetic complications. Methylglyoxal (MGO), one of the major precursors of AGEs, is a highly reactive dicarbonyl compound that plays an important role in the pathogenesis of diabetic nephropathy. This study was designed to evaluate the therapeutic potential of phlorotannin-rich Ecklonia cava extract (ECE) on MGO-induced diabetic nephropathy in in vitro models using mouse glomerular mesangial cells. ECE showed anti-glycation activity via breaking of AGEs-collagen cross-links and inhibition of AGEs formation and AGE-collagen cross-linking formation. The renoprotective effects were determined by assessing intracellular reactive oxygen species (ROS) and MGO accumulation, cell apoptosis, and the Nrf-2/ARE signaling pathway. MGO-induced renal damage, intracellular ROS production level, and MGO-protein adduct accumulation were significantly decreased by pretreating ECE. Moreover, ECE pretreatment exhibited preventive properties against MGO-induced dicarbonyl stress via activation of the Nrf2/ARE signaling pathway and reduction of RAGE protein expression in mouse glomerular mesangial cells. Collectively, these results indicated potential anti-glycation properties and prominent preventive effects of ECE against MGO-induced renal damage. Additionally, ECE may be utilized for the management of AGE-related diabetic nephropathy.